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Updated: Feb 5, 2026

Zygote Microinjection for Creating Gene Cassette Knock-in and Flox Alleles in Mice
Published on: June 24, 2022
A useful gene cassette for conditional knock-down of essential genes by targeted promoter replacement in Mycobacteria
Pauline Texier1, Michèle Coddeville1, Patricia Bordes1
1Laboratoire de Microbiologie et de Génétique Moléculaires, Centre de Biologie Intégrative (CBI), Université de Toulouse, CNRS, UPS, 31062 Toulouse Cedex 09, France.
Abstract:
A direct method to study essential genes is to construct conditional knock-down mutants by replacement of their native promoter by an inducible one. In Mycobacteria, replacement of an essential gene promoter with an anhydrotetracycline inducible one was successfully used but required a multi-step approach. In this work, we describe a gene cassette for the engineering of a conditional knock-down mutant, which allows the one-step targeted replacement of mycobacterial promoters by an anhydrotetracycline-inducible promoter. The functionality of this cassette was successfully tested by engineering conditional clpP and SecA1 mutants of Mycobacterium smegmatis.
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